Method of effecting de-differentiation of a cell

a cell and cell technology, applied in the field of cell dedifferentiation, can solve the problems of affecting the ability of embryonic stem cells to readily differentiate, affecting the differentiation effect of embryonic stem cells, so as to increase the amount or the activity of an err protein, and maintain pluripotency.

Inactive Publication Date: 2011-07-07
AGENCY FOR SCI TECH & RES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0010]In a first aspect the present invention provides a method of effecting de-differentiation of an at least partially differentiated cell or of maintaining pluripotency and / or self-renewing characteristics of an undifferentiated cell. The method includes increasing the amount or the activity of an Err protein, or a functional fragment thereof, in the cell.

Problems solved by technology

It was previously surmised that as stem cells differentiate, they lose their ability to make cell fate decision and become more restricted in their potential.
The ability of embryonic stem cells to readily differentiate furthermore continues to pose a major practical challenge.

Method used

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  • Method of effecting de-differentiation of a cell

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Cell Culture and Transfection

[0091]Mouse ES cells were cultured on gelatin-coated dishes in Dulbecco's modified Eagle medium (DMEM; GIBCO), supplemented with 15% heat-inactivated fetal bovine serum (FBS; GIBCO), 0.055 mM β-mercaptoethanol (GIBCO), 2 mM L-glutamine, 0.1 mM MEM nonessential amino acid, 5,000 units / ml penicillin / streptomycin and 1,000 units / ml of LIF (Chemicon) and passage every 2 ˜3 days. Reprogrammed cells, V6.4 and R1 mouse ES cells were cultured on mitomycin C-treated mouse embryonic fibroblast (MEF) feeders in the same ES cell medium and passage every 2 ˜3 days. MEFs were isolated from 13.5 d.p.c embryos by dissociation with 0.05% trypsin at 37° C. for 10 min and cultured in 15% FBS / DMEM containing 200 μg / ml gentamicin. In this study, we used MEFs within 5 passages to avoid replicative senescence. MEFs from CD1, B6, 129 / B6, Actin-GFP, Actin-GFP / CD1, Pou5fl-GFP / B6 mice have been used for the iPS induction in this study. Transfection of shRNA and over-expression pla...

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Abstract

The invention provides a method of effecting de-differentiation of an at least partially differentiated cell or of maintaining pluripotency and/or self-renewing characteristics of an undifferentiated cell. The method comprises increasing the amount or the activity of an Err protein, or a functional fragment thereof, in the cell.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application makes reference to and claims the benefit of priority of an application for “Methods For Modulating The Differentiation Status Of A Cell” filed on May 6, 2008 with the United States Patent and Trademark Office and there duly assigned the Ser. No. 61 / 050,726. The contents of said application filed on May 6, 2008 is incorporated herein by reference for all purposes, including an incorporation of any element or part of the description, claims or drawings not contained herein and referred to in Rule 20.5(a) of the PCT, pursuant to Rule 4.18 of the PCT.FIELD OF THE INVENTION[0002]The present invention relates to a method of effecting de-differentiation of a cell into a less differentiated cell, including into a pluripotent cell. The method thus allows inter alia forming induced pluripotent stem cells from differentiated somatic cells. The method also allows maintaining pluripotency of an undifferentiated cell.BACKGROUND OF THE...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C12N5/071C12Q1/02G01N33/68C12Q1/25C07H21/00C12N5/0735
CPCC12N5/0696C12N2501/38C12N2510/00C12N2501/603C12N2501/606C12N2501/602
Inventor FENG, BOJIANG, JIANMINGHUI NG, HUCKLUFKIN, THOMASKRAUS, PETRA
Owner AGENCY FOR SCI TECH & RES
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